血吸虫对吡喹酮抗药性的研究XIV日本血吸虫吡喹酮抗性株和敏感株成虫毛蚴和尾蚴对吡喹酮敏感性比较  

Studies on resistance of Schistosoma to praziquantel XIV Experimental comparison of susceptibility to praziquantel between PZQ-resistant isolates and PZQ-susceptible isolates of Schistosoma japonicum in stages of adult worms, miracidia and cercariae

作　　者：李洪军[1] 梁幼生[1] 戴建荣[1] 汪伟[1] 曲国立[1] 李幼子[1] 邢云天[1] 陶永辉[1] 钱科[1] 贾悦[1] 杨振坤[1] 魏剑英[1] 

机构地区：[1]江苏省血吸虫病防治研究所卫生部寄生虫病预防与控制技术重点实验室江苏省寄生虫分子生物学重点实验室,无锡214064

出　　处：《中国血吸虫病防治杂志》2011年第6期611-619,共9页Chinese Journal of Schistosomiasis Control

基　　金：国家科技支撑计划项目(2009BAI78B06);国家自然科学基金(30471516);江苏省自然科学基金(BK2009076);江苏省卫生厅项目(X200912);江苏省预防医学研究课题(Y201031)

摘　　要：目的测定实验室诱导筛选获得的日本血吸虫吡喹酮抗性株成虫、毛蚴和尾蚴对吡喹酮的敏感性变化,为建立吡喹酮敏感性检测/监测技术提供依据。方法以未暴露于吡喹酮的日本血吸虫江苏实验室传代株和现场采集并经实验室传代的湖南株作为吡喹酮敏感株,以实验室传代并经吡喹酮抗性诱导的日本血吸虫江苏抗性诱导株及现场采集并经实验室吡喹酮抗性诱导的湖南抗性诱导株作为吡喹酮抗性株。分别收集各虫株尾蚴感染小鼠,感染35d后分为6组,5个服药组分别一次性灌服剂量为37.5、75、150、300mg/kg和600mg/kg吡喹酮,对照组不给药;用药后14d解剖小鼠,收集小鼠体内成虫,计算虫负荷和减虫率,计算吡喹酮对各虫株的半数有效剂量(ED50值)。取一定量各虫株尾蚴分别暴露于10-5、5×10-6、10-6、5×10-7mol/L和10-7mol/L吡喹酮溶液中,20、40、60、80min和100min后,在解剖镜下观察尾蚴的断尾变化,计算各虫株尾蚴断尾率。取一定量各虫株毛蚴分别暴露于10-5、5×10-6、10-6、5×10-7mol/L和10-7mol/L吡喹酮溶液中,1、3min和5min后,解剖镜下观察毛蚴的形态变化,计算各虫株毛蚴变形率。结果吡喹酮对江苏吡喹酮敏感株和抗性诱导株的ED50值分别为147.7mg/kg和565.5mg/kg,对湖南吡喹酮敏感株和抗性诱导株的ED50值分别为151.8mg/kg和467.2mg/kg。当暴露于10-5mol/L吡喹酮溶液中20min,江苏吡喹酮敏感株尾蚴断尾率为68.8%,吡喹酮抗性诱导株尾蚴断尾率为38.2%,差异有统计学意义(P<0.01);当暴露于10-7mol/L吡喹酮溶液中100min,江苏吡喹酮敏感株尾蚴断尾率为15.9%,吡喹酮抗性诱导株尾蚴断尾率为6.7%,差异有统计学意义(P<0.01)。当暴露于10-5mol/L吡喹酮溶液中20min,湖南吡喹酮敏感株尾蚴断尾率为59.4%,吡喹酮抗性诱导株尾蚴断尾率为54.6%,差异有统计学意义(P<0.05);当暴露于5×10-7mol/L吡喹酮溶液中40min,湖南吡喹酮敏感株尾蚴�Objective To investigate the changes of sensitivity to praziquantel （PZQ） about PZQ-resistant isolates of Schistosoma japonicum established in laboratory by means of the resistance-inducement method during the stages of adult worms, cercariae and miracidia, so as to provide the basis for establishing the sensitivity-detecting technique to praziquantel. Methods A Jiangsu laboratory-maintaining isolate and a Hunan field-collecting isolate of S. japonicum that were never treated with PZQ were as PZQ-susceptible isolates, and two PZQ-induced isolates that were established via drug-treated passage in laboratory were as PZQ-resistant isolates. Mice were infected with S. japonicum cercariae collected from above four isolates each. Thirty-five days after the infection, the mice were divided into 6 groups and administered orally with PZQ at dosages of 0, 37.5, 75, 150, 300 mg/kg and 600 mg/kg, respectively. All the mice were sacrificed two weeks after the treatment, and all the adult worms in the hepatic and portomesenteric veins were recovered and counted. The mean worm burden and reductions were calculated and input into Graphpad Prism 5.0 software, and the PZQ ED50 values of four isolates were calculated by the software. The cercariae of above four isolates were exposed to 10^-5, 5×10^-6, 10^-6, 5×10^-7, 10^-7 mol/L PZQ solutions for 20, 40, 60, 80, 100 min and the changes of tail shedding were observed under a dissecting microscope, then the tail shedding rates of cercariae were calculated. The miracidia of above four isolates were exposed to 5×10^-6, 10^-6, 5×10^-7, 10^-7 mol/L PZQ solutions for 1, 3 and 5 min and the morphological changes were observed under a dissecting microscope, then the morphological change rates of miracidia were calculated. Results The PZQ ED50 values of PZQ-susceptible and PZQ-resistant isolates of Jiangsu were 147.7 mg/kg and 565.5 mg/kg, respectively, and the PZQ ED50 values of PZQ-susceptible and PZQ-resistant isolates of Hunan were 151.8 mg/kg and 467.2 mg/kg, respectively. When

关 键 词：日本血吸虫 吡喹酮 抗药性 成虫 毛蚴 尾蚴 半数有效剂量(ED50) 

分 类 号：R383.24[医药卫生—医学寄生虫学]